The characterization of heterochromatin, the highly repetitive, highly condensed regions of the genome, has lagged behind the characterization of the euchromatic, gene-rich regions. Heterochromatic regions provide important functions, including centromeric activity, which is required for proper distribution of the chromosomes during cell divisions. Using proprietary technology developed by its founders, Dr. Daphne Preuss and colleagues, Chromatin, Inc. will isolate heterochromatic DNA from a variety of crop species, analyze its sequence, and test it for centromere function by constructing an autonomous mini-chromosome vector. This research will provide important information about heterochromatin and centromeres in plants; interestingly, Arabidopsis centromeres contain repeats of a 180- nucleotide sequence, whereas mammalian centromeres contain repeats of a 171-nucleotide sequence. Thus, research on plant heterochromatin may inform studies of mammalian heterochromatin. Furthermore, the ultimate goal of this research is the development of an autonomous mini-chromosome vector, which will be a significant improvement over current transformation methods by allowing the introduction of multiple genes in a controlled regulatory environment. Thus, this research will provide important insight on heterochromatic composition and function and will also provide a valuable tool for research studying the functions of multiple genes. PROPOSED COMMERCIAL APPLICATION: The ultimate goal of this research is the development of an autonomous mini-chromosome vector, which will be a significant improvement over current transformation methods. This vector will allow the introduction of multiple genes, possibly including entire biosynthetic pathways, into plants. Moreover, the autonomy of this vector allows genes to be introduced in a controlled regulatory environment without alteration of the host genome. Possible applications include the synthesis of vitamins or pharmaceuticals in plants and research studying the function of multiple genes in combination.